Supplementary Materials

Supplementary Material for:

Internalization of secreted antigen–targeted antibodies by the neonatal Fc receptor for precision imaging of the androgen receptor axis

Daniel L. J. Thorek, Philip A. Watson, Sang-Gyu Lee, Anson T. Ku, Stylianos Bournazos, Katharina Braun, Kwanghee Kim, Kjell Sjöström, Michael G. Doran, Urpo Lamminmäki, Elmer Santos, Darren Veach, Mesruh Turkekul, Emily Casey, Jason S. Lewis, Diane S. Abou, Marise R. H. van Voss, Peter T. Scardino, Sven-Erik Strand, Mary L. Alpaugh, Howard I. Scher, Hans Lilja,* Steven M. Larson,* David Ulmert*

*Corresponding author. Email: ulmerth{at} (D.U.); liljah{at} (H.L.); larsons{at} (S.M.L.)

Published 30 November 2016, Sci. Transl. Med. 8, 367ra167 (2016)
DOI: 10.1126/scitranslmed.aaf2335

This PDF file includes:

  • Materials and Methods
  • Fig. S1. Anatomical and disease-specific gene expression of candidate targets.
  • Fig. S2. Competition assay.
  • Fig. S3. 89Zr-DFO-11B6 uptake and hK2 expression.
  • Fig. S4. Relative expression of putative prostate markers in PCa cell lines.
  • Fig. S5. 18F-FDHT imaging.
  • Fig. S6. Time-activity curves of LNCaP-AR subcutaneous and orthotopic xenografts.
  • Fig. S7. 89Zr-DFO-11B6 prostate and hK2-specific imaging in transgenic healthy and diseased mice.
  • Fig. S8. Cy5.5-11B6 cellular uptake.
  • Fig. S9. FcRn gene expression.
  • Fig. S10. FcRn-specific transport.
  • Fig. S11. Uptake of pH-responsive dye-labeled 11B6.
  • Fig. S12. Investigation of FcRn-mediated uptake of 11B6 complexed with hK2.
  • Fig. S13. Imaging cross-activation of the AR pathway in LREX′ models.
  • Fig. S14. Accumulation of reengineered anti-PSA antibody.
  • Fig. S15. hK2 production after DHT stimulation.
  • Fig. S16. Intracellular accumulation of 11B6-hK2 in BCa cells.
  • Fig. S17. Quantitation of 89Zr-11B6 uptake in transgenic PCa mice.
  • Fig. S18. Serial PET/CT monitoring 89Zr-11B6 uptake before, during, and after reversible castration by GnRH receptor blockade.
  • Fig. S19. Noninvasive monitoring of AR status with 89Zr-DFO-11B6.
  • Fig. S20. Pathological analysis of sections of GEMM of disease after castration.
  • Fig. S21. Concordance between quantitative ex vivo imaging and protein content.
  • Fig. S22. Comparison of human and murine 11B6.
  • Fig. S23. h11B6 in cynomolgus monkey.
  • Fig. S24. h11B6 immunohistochemistry.
  • Fig. S25. Schematic representation of prostate-specific active hK2 in a genetically engineered KLK2-expressing mouse model.
  • Fig. S26. Genotyping.
  • Table S1. Dissociation rate constants (koff) for m11B6, hu11B6, and DFO-conjugated hu11B6.
  • Table S2. Average association rate constant based on 15 to 18 measurements for each version of 11B6.
  • Table S3. Dissociation rate constants (Kd) for the tested antibodies.
  • Table S4. 89Zr-11B6 biodistribution and the effect of blocking with cold antibody.
  • Table S5. Receptor status of BCa cell lines and secretion of hK2 in response to DHT.
  • Table S6. Data values from PET, bioluminescence, and clinical chemistry measurements.
  • Legend for video S1
  • Reference (51)

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Other Supplementary Material for this manuscript includes the following:

  • Video S1 (.mp4 format). Confocal z-stack through prostate gland of genetically engineered hK2-expressing mouse.